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  3. DiSC3(5)

DiSC3(5) is a fluorescent probe commonly used as a tracer dye to evaluate mitochondrial membrane potential. The excitation/emission wavelength of DiSC3(5) is up to 622/670 nm. DiSC3(5) can inhibit the respiratory system associated with mitochondrial NAD, and the IC50 value is 8 μM. DiSC3(5) in the presence of Na+/K+-ATPase inhibitor ouabain 2 can induce membrane hyperpolarization of Ehrlich ascites tumor cells.

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DiSC3(5) Chemical Structure

DiSC3(5) Chemical Structure

CAS No. : 53213-94-8

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Description

DiSC3(5) is a fluorescent probe commonly used as a tracer dye to evaluate mitochondrial membrane potential. The excitation/emission wavelength of DiSC3(5) is up to 622/670 nm. DiSC3(5) can inhibit the respiratory system associated with mitochondrial NAD, and the IC50 value is 8 μM. DiSC3(5) in the presence of Na+/K+-ATPase inhibitor ouabain 2 can induce membrane hyperpolarization of Ehrlich ascites tumor cells[1][2][3].

In Vitro

1. Preparation of DiSC3(5) membrane staining solution:
1.1 Preparation of DMSO or EtOH reserve solution: The reserve solution should be prepared in DMSO or EtOH at 1-5 mM.
Note 1) : The unused portion of the reserve solution should be stored at -20 ° C. Avoid repeated freezing/thawing cycles.
1.2 Prepare the working solution: Dilute the reserve solution into a suitable buffer, such as serum-free medium, HBSS or PBS, to make a working solution of 1 to 5 uM.
Note 2) : For different cell types and/or experimental conditions, the concentration of the working solution should be determined according to experience. It is recommended to test at concentrations at least in excess of the ten-fold range.
2. Dye the cells into a suspension:
2.1 The suspended cell density in the dye working solution was 1×106/ mL.
2.2 Incubate at 37°C for 2-20 minutes. The culture time depends on the cell type. It is incubated first for 20 minutes and then optimized as needed to obtain even labeling.
2.3 Centrifuge the labeled suspension tubes at 1000 to 1500rpm for 5 minutes.
2.4 Remove the supernatant and gently re-suspend the cells in a preheated (37°C) growth medium.
2.5 Wash twice according to steps 2.3 and 2.4.
3. Staining adherent cells:
3.1 The adherent cells were cultured on a sterile glass cover slide.
3.2 Remove the cover glass from the growth medium and gently drain the excess medium. Place the cover glass in the humidity box.
3.3 Transfer the 100 μL dye working solution to the corner of the cover glass and stir gently until all cells are covered.
3.4 Incubate the cover glass at 37°C for 2-20 minutes. The culture time varies according to cell type. Incubation is started for 20 minutes and then optimized as needed to obtain even labeling.
3.5 Drain the dye working solution and clean the cover glass with growth medium two to three times. For each wash cycle, cover the cells with preheated growth medium, incubate for 5-10 minutes, and then drain the medium.
4. Microscope inspection.
5. Flow cytometry detection:
Dis-labeled cells can be analyzed using conventional FL3 flow cytometry detection channels.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

546.53

Formula

C25H27IN2S2

CAS No.
Appearance

Solid

Color

Blue to dark blue

Emission (Em)

705

Excitation (Ex)

500

SMILES

CCCN1/C(SC2=CC=CC=C12)=C\C=C\C=C\C3=[N+](CCC)C4=CC=CC=C4S3.[I-]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : 16.67 mg/mL (30.50 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.8297 mL 9.1486 mL 18.2973 mL
5 mM 0.3659 mL 1.8297 mL 3.6595 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
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Concentration
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Volume
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Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: 1.25 mg/mL (2.29 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 1.25 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (12.5 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: 1.25 mg/mL (2.29 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 1.25 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (12.5 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.83%

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.8297 mL 9.1486 mL 18.2973 mL 45.7431 mL
5 mM 0.3659 mL 1.8297 mL 3.6595 mL 9.1486 mL
10 mM 0.1830 mL 0.9149 mL 1.8297 mL 4.5743 mL
15 mM 0.1220 mL 0.6099 mL 1.2198 mL 3.0495 mL
20 mM 0.0915 mL 0.4574 mL 0.9149 mL 2.2872 mL
25 mM 0.0732 mL 0.3659 mL 0.7319 mL 1.8297 mL
30 mM 0.0610 mL 0.3050 mL 0.6099 mL 1.5248 mL
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DiSC3(5) Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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